Surface plasmon resonance imaging system and method for measuring molecular interactions

ABSTRACT

A system in an embodiment can comprise an optical assembly, an surface-plasmon-resonance (SPR) light source, and an SPR camera. The optical assembly can comprise a hemispherical prism comprising a top surface configured to support a SPR sensor; and a high numerical aperture (NA) lens located distal from the top surface of the hemispherical prism. The SPR light source can be configured to emit a light beam for SPR imaging. The SPR camera can be configured to capture an SPR image. The SPR sensor further can comprise a surface configured to contact a sample. The high NA lens can be configured to refract the light beam toward the hemispherical prism. The hemispherical prism can be configured to collimate the light beam, as refracted by the high NA lens, toward the SPR sensor. The high NA lens further can be configured to receive and refract the light beam toward the SPR camera, after the light beam is reflected by the surface of the SPR sensor. Other embodiments are disclosed.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of, and claims priority to, U.S.patent application Ser. No. 16/423,733, filed May 28, 2019. U.S. patentapplication Ser. No. 16/423,733 is a continuation of, and claimspriority to, PCT/US19/34087, filed May 27, 2019. PCT/US19/34087 claimspriority to U.S. Provisional Patent Application No. 62/676,983, filedMay 27, 2018. U.S. patent application Ser. No. 16/423,733,PCT/US19/34087, and U.S. Provisional Patent Application No. 62/676,983are incorporated herein by reference in their entirety.

TECHNICAL FIELD

This disclosure relates generally to surface plasmon resonance (SPR)imaging systems, and methods to use such systems, for measuringmolecular interactions.

BACKGROUND

Surface plasmon resonance (SPR) detection using incident light beam is apopular technique for monitoring molecular interactions in real-time.However, traditional SPR devices or systems are not suitable for thestudy of heterogeneity effects naturally occurred in cell populationbecause they either have limited fields of view or are not design forimaging cellular structures or phenotypes that often have randompatterns and structures. Therefore, systems and methods configured tohave a large field of view and a high resolution for measuring molecularinteractions in real time are desired.

BRIEF DESCRIPTION OF THE DRAWINGS

To facilitate further description of the embodiments, the followingdrawings are provided in which:

FIG. 1 illustrates a perspective view of an SPR imaging system,according to an embodiment;

FIG. 2 shows a perspective view of the SPR imaging system, according tothe embodiment in FIG. 1, in which the housing of an optical assembly isomitted;

FIG. 3 shows a side elevational view of the SPR imaging system with thehousing of the optical assembly cut open, according to the embodiment inFIG. 1;

FIG. 4 shows side elevational view of the SPR imaging system with aphase-shift filter, according to the embodiment in FIG. 3;

FIG. 5 shows side elevational view of the SPR imaging system with acolor filter, according to the embodiment in FIG. 3;

FIG. 6 illustrates a cross sectional view of an optical assembly,according to another embodiment;

FIG. 7 shows a side elevational view of an SPR imaging system, accordingto another embodiment;

FIG. 8 shows a top plain view of a sensor translation mount and an SPRsensor of the SPR imaging system, according to the embodiment in FIG. 7;

FIG. 9 shows a partial enlarged view of the SPR imaging system,according the embodiment in FIG. 7;

FIG. 10 shows a side elevational view of an SPR imaging system with thehousing of the optical assembly cut open, according to an embodiment;and

FIG. 11 shows a side elevational view of the SPR imaging system with abeam splitter and a light intensity detector, according to theembodiment in FIG. 10.

For simplicity and clarity of illustration, the drawing figuresillustrate the general manner of construction, and descriptions anddetails of well-known features and techniques may be omitted to avoidunnecessarily obscuring the present disclosure. Additionally, elementsin the drawing figures are not necessarily drawn to scale. For example,the dimensions of some of the elements in the figures may be exaggeratedrelative to other elements to help improve understanding of embodimentsof the present disclosure. The same reference numerals in differentfigures denote the same elements.

The terms “first,” “second,” “third,” “fourth,” and the like in thedescription and in the claims, if any, are used for distinguishingbetween similar elements and not necessarily for describing a particularsequential or chronological order. It is to be understood that the termsso used are interchangeable under appropriate circumstances such thatthe embodiments described herein are, for example, capable of operationin sequences other than those illustrated or otherwise described herein.Furthermore, the terms “include,” and “have,” and any variationsthereof, are intended to cover a non-exclusive inclusion, such that aprocess, method, system, article, device, or apparatus that comprises alist of elements is not necessarily limited to those elements, but mayinclude other elements not expressly listed or inherent to such process,method, system, article, device, or apparatus.

The terms “left,” “right,” “front,” “back,” “top,” “bottom,” “over,”“under,” and the like in the description and in the claims, if any, areused for descriptive purposes and not necessarily for describingpermanent relative positions. It is to be understood that the terms soused are interchangeable under appropriate circumstances such that theembodiments of the apparatus, methods, and/or articles of manufacturedescribed herein are, for example, capable of operation in otherorientations than those illustrated or otherwise described herein.

The terms “couple,” “coupled,” “couples,” “coupling,” and the likeshould be broadly understood and refer to connecting two or moreelements mechanically and/or otherwise. Two or more electrical elementsmay be electrically coupled together, but not be mechanically orotherwise coupled together. Coupling may be for any length of time,e.g., permanent or semi-permanent or only for an instant. “Electricalcoupling” and the like should be broadly understood and includeelectrical coupling of all types. The absence of the word “removably,”“removable,” and the like near the word “coupled,” and the like does notmean that the coupling, etc. in question is or is not removable.

As defined herein, two or more elements are “integral” if they arecomprised of the same piece of material. As defined herein, two or moreelements are “non-integral” if each is comprised of a different piece ofmaterial.

As defined herein, “approximately” can, in some embodiments, mean withinplus or minus ten percent of the stated value. In other embodiments,“approximately” can mean within plus or minus five percent of the statedvalue. In further embodiments, “approximately” can mean within plus orminus three percent of the stated value. In yet other embodiments,“approximately” can mean within plus or minus one percent of the statedvalue.

As defined herein, “real-time” can, in some embodiments, be defined withrespect to operations carried out as soon as practically possible uponoccurrence of a triggering event. A triggering event can include receiptof data necessary to execute a task or to otherwise process information.Because of delays inherent in transmission and/or in computing speeds,the term “real time” encompasses operations that occur in “near” realtime or somewhat delayed from a triggering event. In a number ofembodiments, “real time” can mean real time less a time delay forprocessing (e.g., determining) and/or transmitting data. The particulartime delay can vary depending on the type and/or amount of the data, theprocessing speeds of the hardware, the transmission capability of thecommunication hardware, the transmission distance, etc. However, in manyembodiments, the time delay can be less than approximately one second,five seconds, ten seconds, thirty seconds, one minute, five minutes, tenminutes, or fifteen minutes.

DESCRIPTION OF EXAMPLES OF EMBODIMENTS

Turning to the drawings, FIGS. 1-5 illustrate various views of an SPRimaging system 100, according to an embodiment. In this and otherembodiments, the SPR imaging system 100 can comprise: (a) an opticalassembly 110; (b) an SPR light source 120; (c) an SPR camera 130; (d) abright field light source 140; and/or (e) a bright field camera 150. Inmany embodiments, the SPR imaging system 100 can comprise a high opticalresolution, such as an optical resolution not larger than 3 micrometers(μm), 2 μm, etc. In these and other embodiments, the SPR imaging system100 also can comprise a wide SPR angle, such as an SPR angle rangingfrom 40 to 82 degrees. In some embodiments, the SPR imaging system 100can further comprise a large optical field of view, such as an opticalfield of view as great as 0.1 millimeters-squared (mm²), 1 mm², or 3mm². In many embodiments, the SPR imaging system 100 can be configuredto simultaneously capture or process an SPR image and a bright fieldimage in real time.

In many embodiments, the optical assembly 110 of the SPR imaging system100 can comprise: (a) a hemispherical prism 112 that comprises a planartop surface 113 configured to support a surface-plasmon-resonance (SPR)sensor 170; (b) a high numerical aperture (NA) lens 114; and/or (c) ahousing 116 configured to mount the hemispherical prism 112 and the highNA lens 114, such that the high NA lens 114 is located distal from theplanar top surface 113 of the hemispherical prism 112. In someembodiments, the top surface of the hemispherical prism is not planar oris not entirely planar.

In many embodiments, with the hemispherical prism 112 configured tosupport the SPR sensor 170, no sensor supporting stage is needed, andthe SPR imaging system 100 thus can have fewer heat leaking surfaces andfewer sources of mechanical vibration noise. In many embodiments, thehemispherical prism 112 can comprise a high refractive index, such as arefractive index no less than 1.5. In these and other embodiments, thehigh NA lens 114 can comprise a radius at least 1.5 times greater than aradius of the hemispherical prism 112. In some embodiments, the high NAlens 114 can comprise a high NA value, such as no less than 1.5.

In many embodiments where optical assembly 110 of the SPR imaging system100 comprises the housing 116, the hemispherical prism 112 and the highNA lens 114 can be configured to be firmly coupled to the housing 116 inorder to eliminate any relative movement between the hemispherical prism112 and the high NA lens 114 that can cause mechanical vibration noises.In these and other embodiments, the housing 116 can enclose at least aportion of each of the hemispherical prism 112 and the high NA lens 114for better temperature control. In one such embodiment, the housing 116can enclose the area between the hemispherical prism 112 and the high NAlens 114. In the same or different embodiment, the housing 116 can leavethe planar top surface 113 of the hemispherical prism 112 and the bottomsurface of the high NA lens exposed, as in the exemplary embodimentshown in FIGS. 1 and 3.

In many embodiments, the SPR light source 120 of the SPR imaging system100 can be configured to emit a low-coherent monochromatic light beam121 for SPR imaging toward the high NA lens 114. In many embodiments, anincident angle of the low-coherent monochromatic light beam 121 from theSPR light source 120 towards the high NA lens 114 can be adjustable,such as by one or more additional optical components or by adjusting thelocation or angle of the SPR light source 120. In many embodiments, theSPR camera 130 can be configured to capture an SPR image formed afterthe low-coherent monochromatic light beam 121 is incident upon andreflected by a metal-coated sample contacting surface 171 of the SPRsensor 170.

In many embodiments, the SPR imaging system 100 can comprise the brightfield light source 140 and the bright field camera 150 for bright fieldimaging. In many embodiments, the bright field light source 140 can beconfigured to emit a bright field light beam 141 to illuminate themetal-coated sample contacting surface 171 of the SPR sensor 170, andthe bright field camera 150 can be configured to capture a bright fieldimage of the SPR sensor 170. In some and other embodiments, the brightfield light source 140 can additionally comprise a condenser. As anexample, the condenser can be configured to render a light beam, that isemitted from the bright field light source 140 and originally divergent,into a parallel and/or convergent bright field light beam to illuminatethe SPR sensor 170. In many embodiments, the SPR imaging system 100 canbe configured so that the bright field camera 150 can capture the brightfield image of the SPR sensor 170 simultaneously with the SPR cameracapturing the SPR image.

In many embodiments, the high NA lens 114 can be configured to refractthe low-coherent monochromatic light beam 121 from the SPR light source120 toward the hemispherical prism 112. In the same or differentembodiments, the high NA lens 114 can condition the low-coherentmonochromatic light beam 121 or 122. As an example, the conditioningprovided by the high NA lens 114 can bend the refracting beam, 121 or122, 30-60 degrees from incident beam path, for both beams 121 and 122towards and away from the sensor 170. In many embodiments, thehemispherical prism 112 can be configured to collimate the low-coherentmonochromatic light beam 121, as refracted by the high NA lens 114,toward the SPR sensor 170 that is coupled to the planar top surface 113of the hemispherical prism 112.

In many embodiments, the SPR camera 130 and/or the bright field camera150 can be communicably coupled to a computing device 180 (FIG. 1), suchas a computer or a server, that is configured to: receive and record theSPR image from the SPR camera; receive and record the bright field imagefrom the bright field camera; calibrate the SPR image from the SPRcamera; calibrate the bright field image from the bright field camera;and map the SPR image onto the bright field image for binding analysis.In some embodiments, the computing device 180 (FIG. 1) can be furtherconfigured to automatically perform one or more data processingprocedures, including: (a) normalizing the SPR sensitivity of the sensorsurface per pixel and removing the sensor inhomogeneity by one or morecalibration techniques, such as an SPR profile scan, injection of aknown index standard, a thermal response, or another known method todetermine localized SPR sensor sensitivity; (b) removing sensor drift byleveling the baseline with reference, linear, and non-linearsubtractions; (c) identifying anomalous data of binding behaviors andeliminating artifacts in the data by using Artificial Intelligence(“AI”) to categorize binding behaviors and exclude the data which do notmatch between isotherm analysis and kinetic analysis; and/or (d)identifying one or more sets of data showing model behaviors as themodel data and analyzing the model data to derive a measured result witha certain confidence level.

Furthermore, in many embodiments, the SPR imaging system 100 cancomprise additional optical components, such as one or more lenses, oneor more mirrors, a phase-shift ring 161 (FIG. 4), and/or a color filter162 (FIG. 5), and so forth, for adjusting the colors or directions ofthe light beams and/or the colors or contrast of the bright field imageor the SPR image for SPR imaging, bright field imaging, phase-contrastimaging, and/or fluorescence imaging. For instance, in some embodiments,the phase-shift ring 161 can be configured for phase-contrast imagingwith the bright field light source 140 and the bright field camera 150to increase the contrast of the bright field image, and the phase-shiftring 161 can be located in any suitable place, such as between thehemispherical prism 112 and the high NA lens 114 or between the opticalassembly 110 and the bright field camera 150. In many embodiments, thebright field light source 140 and the bright field camera 150 can befurther configured for fluorescence imaging with one of: (a) the brightfield light beam 141 (see also, 341 (FIG. 7)) emitted by the brightfield light source 140 comprising a colored light; or (b) the brightfield light beam 141 emitted by the bright field light source 140comprising a white light and the color filter 162 located at one of:between the bright field light source 140 and the SPR sensor 170 or infront of bright field camera 150, and configured to change one or morecolors of bright field light beam 141/142 (see also, 341/342 (FIG. 7)).

Turning to the drawings, FIG. 6 illustrates an optical assembly 200,according to an embodiment. In many embodiments, the optical assembly200 can comprise a hemispherical prism 212, a high NA lens 214, and/or ahousing 216. In many embodiments, the hemispherical prism 212 cancomprise a planar top surface 213 configured to support an SPR sensor270 with a metal-coated sample contacting surface 271. In someembodiments, the top surface of the hemispherical prism is not planar oris not entirely planar. In many embodiments, the housing 216 can beconfigured to mount the hemispherical prism 212 and the high NA lens214, such that the high NA lens 214 is located distal from the planartop surface 213 of the hemispherical prism 212. In many embodiments, anSPR imaging system adopting the optical assembly 200 can comprise: ahigh optical resolution, such as an optical resolution not larger than 3μm, 1 μm, etc.; a wide SPR angle, such as an SPR angle ranging from 40to 82 degrees; and/or a large optical field of view, such as an opticalfield of view as great as 0.1 mm², 1 mm², or 3 mm². In many embodiments,the optical assembly 200 can be used for simultaneous SPR and brightfield imaging.

In many embodiments, the hemispherical prism 212 can comprise a highrefractive index, such as a refractive index no less than 1.5. In theseand other embodiments, the high NA lens 214 can comprise a radius atleast 1.5 times greater than a radius of the hemispherical prism 212. Insome embodiments, the high NA lens 214 can comprise a high NA value,such as no less than 1.5. In many embodiments, the hemispherical prism212 and the high NA lens 214 can be configured to be firmly coupled tothe housing 216 in order to eliminate any relative movement between thehemispherical prism 212 and the high NA lens 214 that can causemechanical vibration noise. In these and other embodiments, the housing216 can enclose at least a portion of each of the hemispherical prism212 and the high NA lens 114 for better temperature control, such asenclosing the area between the hemispherical prism 212 and the high NAlens 214.

Turning to the drawings, FIGS. 7-9 illustrate various views of an SPRimaging system 300, according to another embodiment. In this and otherembodiments, the SPR imaging system 300 can comprise: (a) an opticalassembly 310; (b) an SPR light source 320; (c) an SPR camera 330; (d) abright field light source 340; (e) a bright field camera 350; (f) athermoelectric device 318; (g) a microfluidic device 372; and/or (h) asensor translation mount 373 (FIGS. 8-9). In many embodiments, theoptical assembly 310 can comprise a hemispherical prism 312, a high NAlens 314, a thermoelectric device 318, and/or a housing 316. In manyembodiments, the hemispherical prism 312, the high NA lens 314, and thehousing 316 can be similar to the aforementioned hemispherical prism(112 (FIG. 1-5) or 212 (FIG. 6)), the high NA lens (114 (FIG. 1-5) or214 (FIG. 6)), and/or the housing (116 (FIG. 1-5) or 216 (FIG. 6)),respectively.

In many embodiments, the thermoelectric device 318 can be configured tocontrol the temperature of the SPR sensor 370, such as maintaining thetemperature fluctuation of the SPR sensor 370 within 0.016 degreesCelsius (° C.), in order to avoid noises such as baseline shift in theSPR response signal or change in SPR angle. In some embodiments, thethermoelectric device 318 can be located within the optical assembly310, such as near the top surface of the hemispherical prism 312, and/orpartially or entirely surrounding the hemispherical prism 312, in orderto maintain the temperature of the hemispherical prism 312 and, in turn,maintain the temperature of the SPR sensor 370.

In many embodiments, the microfluidic device 372 can be mountable on theSPR sensor 370 and configured to deliver a buffer solution with one ormore ligand samples 363 onto the metal-coated sample contacting surfaceof the SPR sensor 370. In some embodiments, the microfluidic device 372also can comprise: a pump with buffer exchange and degas capabilityconfigured to control a flow of the buffer solution; and/or anauto-sampler configured to place one or more ligand samples onto themetal-coated sample contacting surface of the SPR sensor 370.

In many embodiments, the SPR imaging system 300 can comprise the sensortranslation mount 373 to monitor more cell population on a single SPRsensor 370. In many embodiments, the sensor translation mount 373 can bemountable on a planar top surface 313 of the hemispherical prism 312 andconfigured to hold the SPR sensor 370 and translate or move the SPRsensor 370, by an x adjust arm 3731 (FIGS. 8-9) and a y adjust arm 3732(FIG. 8), on the planar top surface 313 of hemispherical prism 312,e.g., within a few millimeters on the planar top surface 313, to expandthe measuring area on the SPR sensor 370, such as a measuring area asmuch as 4 times the area of the optical field of view of the SPR sensor370. In many embodiments, a thin layer of matching oil (not shown) alsocan be applied between the SPR sensor 370 and the planar top surface 313of the hemispherical prism 312 to eliminate any interface effect andprovide lubricant for sensor translation. In some embodiments, the topsurface of the hemispherical prism 312 is not planar or is not entirelyplanar.

Turning to the drawings, FIGS. 10-11 show an SPR imaging system 400,according to an embodiment. In many embodiments, the SPR imaging system400 can comprise a motorized frame 423 configured to change an incidentangle ø of a low-coherent monochromatic light beam 421 relative to anoptical assembly 410. As known in the art, a typical intensity-angleprofile curve comprises a dip at the SPR angle, and an angle spreadbetween the two steep slope regions near the dip is where a small angleshift can cause significant change in an intensity of SPR light shown inan SPR image; thus, measuring at the angle spread provides the highestsensitivity for SPR measurement. In many embodiments, the SPR imagingsystem 400 can be configured to automatically scan and/or adjust theangle ø of an incident low-coherent monochromatic light beam 421 toprovide a wide range of angle adjustment, e.g., 40-82 degrees. The angleadjustment allows the detector to optimize the sensitivity in SPRmeasurement. For instance, in some embodiments, the motorized frame 423configured can be configured to move the SPR light source 420 verticallyor in one linear direction only to adjust the light path and then theincident angle Π of the low-coherent monochromatic light beam 421.

Additionally, as known in the art, the stability of an SPR light source,such as an SPR light source 420, can have a significant effect on SPRresponse signal measurement, and the measured reflectivity intensityI_(m) is proportional to the incident light intensity I_(o) times afunction a(n) of sensor surface property and refractive index at and/ornear its surface. That is, I_(m)=a(n)×I_(o). To minimize this dependencyof I_(o), a normalized SPR signal I_(spr) can be used:

I _(spr) =I _(m) /I _(o) =a(n), independent of I _(o).

Therefore, in many embodiments, the SPR imaging system 400 can furthercomprise a light intensity detector 425 (FIG. 11) configured to monitorand record the intensity of the low-coherent monochromatic light beam421 for intensity normalization and noise reduction. In these and otherembodiments, the SPR imaging system 400 also can comprise a beamsplitter 424 (FIG. 11), such as a 50% beam splitter, located in thelight path of incident low-coherent monochromatic light beam 421 tosplit the incident low-coherent monochromatic light beam 421 into twoportions. In many embodiments, with the light intensity detector 425and/or the beam splitter 424, the SPR imaging system 400 can beconfigured to automatically detect and/or maintain the intensity of thelow-coherent monochromatic light beam 421.

In an embodiment, a system can comprise an optical assembly (e.g., 110(FIGS. 1-5), 200 (FIG. 6), 310 (FIG. 9), or 410 (FIGS. 10-11)), an SPRlight source (e.g., 120 (FIGS. 1-5), 320 (FIG. 7), or 420 (FIGS.10-11)), and/or an SPR camera (e.g., 130 (FIGS. 1-5), 330 (FIG. 7), or430 (FIGS. 10-11)). The optical assembly in this and other embodimentscan comprise a hemispherical prism (e.g., 112 (FIGS. 1-5), 212 (FIG. 6),312 (FIGS. 7 & 9), or 412 (FIGS. 10-11)) that comprises a top surfaceconfigured to support a surface-plasmon-resonance (SPR) sensor (e.g.,170 (FIGS. 1-5), 270 (FIG. 6), 370 (FIG. 7-9), or 470 (FIGS. 10-11)); ahigh numerical aperture (NA) lens (e.g., 114 (FIGS. 1-5), 214 (FIG. 6),314 (FIGS. 7 & 9), or 414 (FIGS. 10-11)); and a housing (e.g., 116(FIGS. 1-5), 216 (FIG. 6), 316 (FIGS. 7 & 9), or 416 (FIGS. 10-11))configured to mount the hemispherical prism and the high NA lens suchthat the high NA lens is located distal from the top surface of thehemispherical prism. The SPR light source in this and other embodimentscan be configured to emit a low-coherent monochromatic light beam (e.g.,121 (FIGS. 1-5), 221 (FIG. 6), 321 (FIG. 7), or 421 (FIGS. 10-11)) forSPR imaging toward the high NA lens. The SPR camera in this and otherembodiments can be configured to capture an SPR image formed after thelow-coherent monochromatic light beam (e.g., 122 (FIGS. 1-5), 222 (FIG.6), or 322 (FIG. 7)) is incident upon and reflected by a metal-coatedsample contacting surface (e.g., 171 (FIG. 1-5), or 271 (FIG. 6)) of theSPR sensor. Additionally, in this and other embodiments, the high NAlens can be configured to refract the low-coherent monochromatic lightbeam from the SPR light source toward the hemispherical prism; and thehemispherical prism can be configured to collimate the low-coherentmonochromatic light beam, as refracted by the high NA lens, toward theSPR sensor.

In another embodiment, a method for surface-plasmon-resonance (SPR)imaging can comprise: (a) coupling an SPR sensor to an optical assembly;(b) placing one or more ligand samples on a metal-coated samplecontacting surface of the SPR sensor; (c) emitting a low-coherentmonochromatic light beam from an SPR light source toward the high NAlens; and/or (d) capturing an SPR image by an SPR camera. In this andother embodiments, the optical assembly can comprise a hemisphericalprism comprising a top surface configured to support the SPR sensor; ahigh numerical aperture (NA) lens; and/or a housing configured to mountthe hemispherical prism and the high NA lens such that the high NA lensis located distal from the top surface of the hemispherical prism. Inaddition, in many embodiments, the high NA lens can be configured torefract the low-coherent monochromatic light beam from the SPR lightsource toward the hemispherical prism. Further, the hemispherical prismin these and other embodiments can be configured to collimate thelow-coherent monochromatic light beam, as refracted by the high NA lens,toward the SPR sensor. Moreover, in these and other embodiments, the SPRimage can be formed after the low-coherent monochromatic light beam isincident upon and reflected by the metal-coated sample contactingsurface of the SPR sensor.

In yet another embodiment, an optical assembly for surface plasmonresonance (SPR) imaging can comprise: (a) a hemispherical prismcomprising a high refractive index no less than 1.5 and a planar topsurface configured to support an SPR sensor; (b) a high numericalaperture (NA) lens comprising a radius at least 1.5 times greater than aradius of the hemispherical prism; (c) a housing configured to mount thehemispherical prism and the high NA lens such that the lens is locateddistal from the planar top surface of the hemispherical prism; (d) ahigh optical resolution, not larger than 3 μm; (e) a wide SPR angle,ranging from 40 to 82 degrees; and (f) a large optical field of view,not less than 0.1 mm². In this and other embodiments, the high NA lenscan be configured to refract a light toward the hemispherical prism;and/or the hemispherical prism can be configured to collimate the light,as refracted by the high NA lens, toward the SPR sensor.

In some embodiments, a system can comprise an optical assembly, an SPRlight source, and/or an SPR camera. The optical assembly can comprise ahemispherical prism that comprises a top surface configured to support asurface-plasmon-resonance (SPR) sensor; and a high numerical aperture(NA) lens located distal from the top surface of the hemisphericalprism. The SPR light source can be configured to emit a light beam forSPR imaging. The SPR camera can be configured to capture an SPR image.In these and other embodiments, the SPR sensor further can comprise asurface configured to contact a sample. The high NA lens can beconfigured to refract the light beam toward the hemispherical prism. Thehemispherical prism can be configured to collimate the light beam, asrefracted by the high NA lens, toward the SPR sensor. The high NA lensfurther can be configured to receive and refract the light beam towardthe SPR camera, after the light beam is reflected by the surface of theSPR sensor.

In a number of embodiments, a method for surface-plasmon-resonance (SPR)imaging can comprise: (a) coupling an SPR sensor to an optical assembly;(b) placing one or more ligand samples on a surface of the SPR sensor;(c) emitting a light beam from an SPR light source; and/or (d) capturingan SPR image by an SPR camera. In these and other embodiments, theoptical assembly can comprise a hemispherical prism comprising a topsurface configured to support the SPR sensor; and a high numericalaperture (NA) lens located distal from the top surface of thehemispherical prism. In addition, in some embodiments, the high NA lenscan be configured to refract the light beam from the SPR light sourcetoward the hemispherical prism. The hemispherical prism can beconfigured to collimate the light beam, as refracted by the high NAlens, toward the SPR sensor. The SPR image can be formed after the lightbeam is incident upon and reflected by the surface of the SPR sensor.The high NA lens further can be configured to receive and refract thelight beam toward the SPR camera, after the light beam is reflected bythe surface of the SPR sensor.

In certain embodiments, an optical assembly for surface plasmonresonance (SPR) imaging can comprise: (a) a hemispherical prismcomprising a high refractive index no less than 1.5 and a planar topsurface configured to support an SPR sensor; (b) a high numericalaperture (NA) lens comprising a radius at least 1.5 times greater than aradius of the hemispherical prism; (c) a high optical resolution, notlarger than 3 μm; (d) a wide SPR angle, ranging from 40 to 82 degrees;and (e) a large optical field of view, not less than 0.1 mm². In theseand other embodiments, the high NA lens can be located distal from thetop surface of the hemispherical prism and configured to refract a lighttoward the hemispherical prism. The hemispherical prism can beconfigured to: (a) collimate the light, as refracted by the high NAlens, toward the SPR sensor; and (b) receive and refract the light,reflected by the SPR sensor, toward the high NA lens. The high NA lensfurther can be configured to receive the light, as refracted from thehemispherical prism.

Although systems and methods for SPR imaging and/or simultaneous brightlight imaging have been described with reference to specificembodiments, it will be understood by those skilled in the art thatvarious changes may be made without departing from the spirit or scopeof the disclosure. Accordingly, the disclosure of embodiments isintended to be illustrative of the scope of the disclosure and is notintended to be limiting. It is intended that the scope of the disclosureshall be limited only to the extent required by the appended claims. Forexample, to one of ordinary skill in the art, it will be readilyapparent that any components of the SPR imaging system(s) and/or opticalassemblies, as well as the steps to use the SPR imaging system(s) and/oroptical assemblies, may be modified, and that the foregoing discussionof certain of these embodiments does not necessarily represent acomplete description of all possible embodiments.

Replacement of one or more claimed elements constitutes reconstructionand not repair. Additionally, benefits, other advantages, and solutionsto problems have been described with regard to specific embodiments. Thebenefits, advantages, solutions to problems, and any element or elementsthat may cause any benefit, advantage, or solution to occur or becomemore pronounced, however, are not to be construed as critical, required,or essential features or elements of any or all of the claims, unlesssuch benefits, advantages, solutions, or elements are stated in suchclaim.

Moreover, embodiments and limitations disclosed herein are not dedicatedto the public under the doctrine of dedication if the embodiments and/orlimitations: (1) are not expressly claimed in the claims; and (2) are orare potentially equivalents of express elements and/or limitations inthe claims under the doctrine of equivalents.

What is claimed is:
 1. A system comprising: an optical assemblycomprising: a hemispherical prism comprising a top surface configured tosupport a surface-plasmon-resonance (SPR) sensor; and a high numericalaperture (NA) lens located distal from the top surface of thehemispherical prism; an SPR light source configured to emit a light beamfor SPR imaging; and an SPR camera configured to capture an SPR image,wherein: the SPR sensor further comprises a surface configured tocontact a sample; the high NA lens is configured to refract the lightbeam toward the hemispherical prism; the hemispherical prism isconfigured to collimate the light beam, as refracted by the high NAlens, toward the SPR sensor; and the high NA lens is further configuredto receive and refract the light beam toward the SPR camera, after thelight beam is reflected by the surface of the SPR sensor.
 2. The systemof claim 1, wherein: the hemispherical prism further comprises arefractive index no less than 1.5; and the high NA lens furthercomprises a radius at least 1.5 times greater than a radius of thehemispherical prism.
 3. The system of claim 2, wherein the opticalassembly further comprises: a high optical resolution, not larger than 3μm; a wide SPR angle, ranging from 40 to 82 degrees; and a large opticalfield of view, not less than 0.1 mm².
 4. The system of claim 1, whereinthe optical assembly further comprises a thermoelectric deviceconfigured to control a temperature of the SPR sensor coupled to thehemispherical prism.
 5. The system of claim 1, wherein an incident angleof the light beam from the SPR light source towards the high NA lens isadjustable.
 6. The system of claim 1 further comprising a bright fieldlight source and a bright field camera for bright field imaging,wherein: the bright field light source is configured to emit a brightfield light beam to illuminate the surface of the SPR sensor; and thebright field camera is configured to capture a bright field image of theSPR sensor simultaneously with the SPR camera capturing the SPR image.7. The system of claim 6 further comprising a phase-shift ring locatedbetween the hemispherical prism and the high NA lens and configured forphase-contrast imaging with the bright field light source and the brightfield camera.
 8. The system of claim 6, wherein the bright field lightsource and the bright field camera are further configured forfluorescence imaging with one of: the bright field light beam emitted bythe bright field light source comprising a colored light; or the brightfield light beam emitted by the bright field light source comprising awhite light and a color filter located at one of: between the brightfield light source and the SPR sensor or in front of the bright fieldcamera, the color filter configured to change one or more colors of thebright field light beam.
 9. The system of claim 6, wherein: the SPRcamera and the bright field camera are communicably coupled to acomputing system; the SPR camera is further configured to transmit theSPR image to the computing system; the bright field camera is furtherconfigured to transmit the bright field image to the computing system;and the computing system is configured to: receive and record the SPRimage transmitted from the SPR camera; receive and record the brightfield image transmitted from the bright field camera; calibrate the SPRimage received from the SPR camera; calibrate the bright field imagereceived from the bright field camera; and map the SPR image onto thebright field image for binding analysis.
 10. The system of claim 1further comprising a light intensity detector configured to monitor andrecord an intensity of the light beam for intensity normalization andnoise reduction.
 11. A method for surface-plasmon-resonance (SPR)imaging comprising: coupling an SPR sensor to an optical assembly, theoptical assembly comprising: a hemispherical prism comprising a topsurface configured to support the SPR sensor; and a high numericalaperture (NA) lens located distal from the top surface of thehemispherical prism; placing one or more ligand samples on a surface ofthe SPR sensor; emitting a light beam from an SPR light source; andcapturing an SPR image by an SPR camera, wherein: the high NA lens isconfigured to refract the light beam from the SPR light source towardthe hemispherical prism; the hemispherical prism is configured tocollimate the light beam, as refracted by the high NA lens, toward theSPR sensor; the SPR image is formed after the light beam is incidentupon and reflected by the surface of the SPR sensor; and the high NAlens is further configured to receive and refract the light beam towardthe SPR camera, after the light beam is reflected by the surface of theSPR sensor.
 12. The method of claim 11, wherein: the hemispherical prismfurther comprises a refractive index no less than 1.5; and the high NAlens further comprises a radius at least 1.5 times greater than a radiusof the hemispherical prism.
 13. The method of claim 12, wherein theoptical assembly further comprises: a high optical resolution, notlarger than 3 μm; a wide SPR angle, ranging from 40 to 82 degrees; and alarge optical field of view, not less than 0.1 mm².
 14. The method ofclaim 11, wherein the optical assembly further comprises athermoelectric device configured to control a temperature of the SPRsensor coupled to the hemispherical prism.
 15. The method of claim 11further comprising adjusting an incident angle of the light beam fromthe SPR light source towards the high NA lens to allow scanning an SPRangle profile and to optimize a detection sensitivity.
 16. The method ofclaim 11 further comprising: emitting a bright field light beam from abright field light source to illuminate the surface of the SPR sensor;and capturing a bright field image of the SPR sensor by a bright fieldcamera simultaneously with capturing the SPR image by the SPR camera.17. The method of claim 16 further comprising: placing a phase-shiftring between the hemispherical prism and the high NA lens and configuredfor phase-contrast imaging with the bright field light source and thebright field camera.
 18. The method of claim 16, wherein the brightfield light source and the bright field camera are further configuredfor fluorescence imaging with one of: changing a color of the brightfield light beam emitted by the bright field light source; or placing acolor filter at one of: between the bright field light source and theSPR sensor or in front of the bright field camera, the color filterconfigured to change one or more colors of the bright field light beam.19. The method of claim 16 further comprising: transmitting the SPRimage from the SPR camera to a computing system; and transmitting thebright field image from the bright field camera to the computing system,wherein: the computing system is configured to: receive and record theSPR image transmitted from the SPR camera; receive and record the brightfield image transmitted from the bright field camera; calibrate the SPRimage received from the SPR camera; calibrate the bright field imagereceived from the bright field camera; and map the SPR image onto thebright field image for binding analysis.
 20. The method of claim 11further comprising providing a light intensity detector for intensitynormalization and noise reduction, the light intensity detectorconfigured to monitor and record an intensity of the light beam.
 21. Anoptical assembly for surface plasmon resonance (SPR) imaging comprising:a hemispherical prism comprising a high refractive index no less than1.5 and a top surface configured to support an SPR sensor; a highnumerical aperture (NA) lens comprising a radius at least 1.5 timesgreater than a radius of the hemispherical prism; a high opticalresolution, not larger than 3 μm; a wide SPR angle, ranging from 40 to82 degrees; and a large optical field of view, not less than 0.1 mm²,wherein: the high NA lens is located distal from the top surface of thehemispherical prism and configured to refract a light toward thehemispherical prism; the hemispherical prism is configured to: (a)collimate the light, as refracted by the high NA lens, toward the SPRsensor, and (b) receive and refract the light, reflected by the SPRsensor, toward the high NA lens; and the high NA lens is furtherconfigured to receive the light, as refracted from the hemisphericalprism.
 22. The optical assembly of claim 21 further comprising athermoelectric device configured to control a temperature of the SPRsensor coupled to the hemispherical prism.